We narrowed to 749 results for: Tro;
-
TypeBlog PostPublishedApril 14, 2016, 2:30 p.m....later, it was shown that putting the operon under a strong promoter results in high R-body expression (Schrallhammer...bodies could also perform as actuators in microelectromechanical systems (MEMS) devices. They could be functionalized...of armor to protect their host from predation (Petroni et al 2000). With a better understanding of how... Plasmids from This Publication at Addgene 4. Petroni, G., Spring, S., Schleifer, K.-H., Verni, F. & ...
-
Top Requested Lentivirus and AAV of 2016
TypeBlog PostPublishedJan. 6, 2017, 3:56 p.m....unique editing tasks, classic Cas9 nuclease, which introduces double stranded DNA breaks, can be expressed ...Gαi-coupled) DREADD fused to mCherry, all under the control of the human synapsin promoter. Purpose of this...mCherry can be used to induce neuronal silencing in vitro and in vivo (Krashes et al., 2011). The various ...Rumpel S. Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, ... -
Plasmids 101: Methylation and Restriction Enzymes
TypeBlog PostPublishedJune 30, 2016, 2:30 p.m....digest as depicted in Lane 2. Controlling methylation Finally, you can control methylation by altering your...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which... with the Latest Cloning Techniques Learn to Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis... -
Antibodies 101: Secondary Antibodies
TypeBlog PostPublishedDec. 21, 2021, 2:15 p.m....antibodies. The introduction of antibody fragments, such as nanobodies, has introduced new options for...Antibodies 101: Introduction to Immunofluorescence Antibodies 101: ELISAs Antibodies 101: Introduction to Antibodies... -
Antibodies 101: Polyclonal Antibodies
TypeBlog PostPublishedJuly 14, 2021, 1:15 p.m....antibody, you’ll want to use the same positive control across experiments to be aware of signal strength...you’ll want to be sure to choose the appropriate controls to be sure the staining you are seeing is real...real. It’s important to include a negative control using serum from the same species your polyclonal antibody...antibody was generated in or a polyclonal isotype control, to account for nonspecific binding. Now that you... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog PostPublishedMay 11, 2021, 1:15 p.m....localization. Surprisingly our observations at the electron microscope showed us that, in monkey neurons, ...of the haemagglutinin of the influenza virus, introduced to the field of fusion proteins more than thirty... of interest the membrane. Figure 1: Electron microscope images showing ultrastructural localization...protein of interest, an alternative could be bicistronic expression of the fluorescent protein with the... -
Plasmids 101: CcdB - The Toxic Key to Efficient Cloning
TypeBlog PostPublishedDec. 8, 2016, 3:30 p.m....the insert. Gateway® technology (developed by InvitrogenTM) is essentially a more modern version of this...teachline.ls.huji.ac.il/72682/tutorials/gateway/introduction.html demonstrating the principal behind the “swapping...CcdB-resistant strain is ccdB Survival™ from InvitrogenTM. Using either DB3.1 or ccdB SurvivalTM, one ...PubMed PMID: 1324324. 6. Tam, JE. and Kline, BC. “Control of the ccd operon in plasmid F.” J Bacteriol. 1989... -
Plasmids 101: Cre-lox
TypeBlog PostPublishedJan. 13, 2015, 3:47 p.m....AAV (AAV-pgk-Cre) has been used to successfully introduce Cre into cells of interest. What if I need two...of using loxP sites is the inability to tightly control which loxP sites recombine if more than two are...allowing for two separate recombination events to be controlled in parallel. Have you used the Cre-lox system...PubMed Central PMCID: PMC49604. 4. Independent control of immunoglobulin switch recombination at individual... -
Split Fluorescent Proteins for Studying Protein-Protein Interactions
TypeBlog PostPublishedMay 3, 2022, 1:15 p.m....live cells. Controls When designing experiments, it’s important to include proper controls to assess the...quantitative readout of the interaction in vivo or in vitro, and can be used for measuring protein expression... p53 tetramerization in response to DNA damage controls the p53 response. Gaglia G, Lahav G. Mol Syst ... -
Viral Vectors 101: Chemogenetics
TypeBlog PostPublishedJuly 11, 2023, 1:15 p.m....chemogenetics? Chemogenetics relies on genetically introducing engineered receptors that are only activated ...Expression Using chemogenetics requires successfully introducing receptors to target cells. Targeted and specific...mitigated by using low doses and running appropriate controls (Jendryka et al., 2019), they have resulted in...channel, the PSAM acts as a gating mechanism to control ion flux. The system allows you to directly manipulate... -
No Llamas Required - Synthetic Nanobodies Against Membrane Proteins
TypeBlog PostPublishedJune 18, 2020, 1:15 p.m.... proteins. The Seeger lab developed a robust in-vitro protocol that enables rapid selection of synthetic...sybodies that hypothetically contain ones that bind strongly to your target protein. The Seeger lab designed...selection step involves using a commercial kit for in-vitro translation of the mRNA libraries. Each mRNA molecule...enrichment step for identifying sybodies that bind strongly and specifically to the protein of interest. The... -
Of Myc and Men
TypeBlog PostPublishedJan. 19, 2023, 2:15 p.m....of interest to better study them in vivo and in vitro, especially in the mid-late 20th century with the...reticulum localization, Munro and Pelham casually introduced the Myc-tag (Munro, et al. 1986; Munro and Pelham... and the Myc-tag today Since Munro and Pelham introduced the Myc tag to the world, it has become one of...this antibody works and allow users to better troubleshoot and design experiments. It turns out that even... -
Plasmids 101: Degron Tags
TypeBlog PostPublishedMarch 14, 2024, 1:15 p.m....advantages degron tags can offer over other protein control methods and compare the commonly available types...etc. Firstly, they are reversible mechanisms of control. Protein degradation can be induced by the addition... Tsien RY, Lin MZ. Tunable and reversible drug control of protein production via a self-excising degron... Susanna Stroik... -
Truncated gRNAs for Regulating Gene Expression
TypeBlog PostPublishedJan. 10, 2017, 3:37 p.m....can be exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast...necessary gRNAs, but it would not be possible to control which gRNA becomes complexed with which variant...represents a straightforward means of exerting tight control over Cas9 nuclease activity while eliminating the...guides targeting one set of genetic loci can be introduced together with truncated gRNAs targeting a different... -
Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards
TypeBlog PostPublishedMay 22, 2015, 4:58 p.m....experiments attempting to build gene drives in tropical mosquito vectors of diseases such as malaria and...then, using multiple confinement strategies is strongly advised. The effects are multiplicative, and there.... Site-specific selfish genes as tools for the control and genetic engineering of natural populations....Fitz-Earle, M., Holm, D. G. & Suzuki, D. T. Genetic Control of Insect Populations: I. Cage Studies of Chromosome... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....and would avoid genomic off-targets or indels introduced through non-homologous end joining (NHEJ). Designing...recapitulate or rescue known pathogenic alleles, or introduce a premature stop codon to render an RNA nonfunctional...also adjusted the structure of the guide RNA by introducing a C opposite the target A to be edited. This ...off-target edits around the targeted nucleotide. Introduction of guanine-mismatches in the guide across from... -
Negotiating Work and Life: How to Find the Joy
TypeBlog PostPublishedMarch 20, 2014, 3:04 p.m....for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood. Work/life balance....possible to bring all aspects of our lives under control? Balance is a myth and should not even be the real... don’t have children, so my life will be under control.” Think again. Maybe you’ll need to care for an...to do most tasks – try to resist the impulse to control by insisting things always be done your way. Be... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog PostPublishedJuly 7, 2020, 1:15 p.m.... Martin et al., 2018). The ACE reporter is a bicistronic construct that consists of a mutated mCherry ...eGFP. To create the inactive mCherry, the lab introduced a 43 base-pair insertion that disrupts fluorescence...use across different cell types and systems. To introduce flexibility, the Dow lab created a base editing... optimized cytidine base editor. The lab then introduced an sgRNA targeting the GFPACG to the cells. As... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog PostPublishedJuly 13, 2023, 1:15 p.m....protocols therefore differ from library to library. We strongly recommend following the protocols provided by ...a low multiplicity of infection (MOI). MOI is controlled by viral titer, so you’ll need to measure your...Addgene uses a physical titer (via ddPCR) to quality control our viral preps, we recommend determining the functional...transduction protocol you can follow and a troubleshooting blog should you run into any issues. Keep ... -
Supporting Reproducibility with a Connected ELN
TypeBlog PostPublishedJan. 17, 2019, 1:38 p.m....founder of Research Space. There are many types of electronic lab notebooks (ELNs), each with its own pros ... virtue of liberating data from paper into an electronic environment and hence making it searchable and... of Research Space, which provides the RSpace electronic lab notebook. Rory is based in Cambridge, Massachusetts...on the Addgene blog Find tips on choosing an electronic lab notebook Watch a livestream of the Minisymposium...